Difference between revisions of "Team:WashU StLouis/Proof"
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After analyzing the results of our absorbance data of electron donors, it was decided to go a step further to ensure that these construct produced usable electron donors. A biotin assay was found that offered an indirect way of measuring the amount of reduced electron donors present. Biotin, commonly know as vitamin B7, is produced by a pathway that uses reduced flavodoxin/ferredoxin,specifically the SAM pathway that turns dethiobiotin to biotin (see Figure 1). | After analyzing the results of our absorbance data of electron donors, it was decided to go a step further to ensure that these construct produced usable electron donors. A biotin assay was found that offered an indirect way of measuring the amount of reduced electron donors present. Biotin, commonly know as vitamin B7, is produced by a pathway that uses reduced flavodoxin/ferredoxin,specifically the SAM pathway that turns dethiobiotin to biotin (see Figure 1). | ||
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| − | + | <img src = "https://static.igem.org/mediawiki/2016/b/b2/T--WashU_StLouis--BiotinSynthesis.gif" style="display: block; margin: 0 auto; width: 400px;"> | |
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| + | By measuring the amount of biotin in the new cells, and recognizing the 2:1 stoichiometric relationship between reduced electron donors and biotin molecules, the amount of electron donors present and used can be indirectly measured. Results showed that __________, as seen below: | ||
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| + | <img src = "https://static.igem.org/mediawiki/2016/c/c7/T--WashU_StLouis--BiotinDH10Bgraph.png" style="width : 40vw;display: block; margin: 0 auto;"> | ||
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| + | <img src = "https://static.igem.org/mediawiki/2016/b/be/T--WashU_StLouis--BiotinYalegraph2.png" style="width : 50vw;display: block; margin: 0 auto;"> | ||
| + | <!--<img src = "https://static.igem.org/mediawiki/2016/c/c5/T--WashU_StLouis--BiotinYalegraph.png" style="width : 700px">--> | ||
Revision as of 17:36, 17 October 2016
Proof of Concept
Showing that our stuff could make things
After identifying the genes and constructing the electron donor overproducing plasmids, a proper assay was necessary in order to see if our new cells produced usable forms of both electron donors. Not only was it shown that the products were overproduced, but also significant data supported the claim that these products were usable in protein synthesis.
After analyzing the results of our absorbance data of electron donors, it was decided to go a step further to ensure that these construct produced usable electron donors. A biotin assay was found that offered an indirect way of measuring the amount of reduced electron donors present. Biotin, commonly know as vitamin B7, is produced by a pathway that uses reduced flavodoxin/ferredoxin,specifically the SAM pathway that turns dethiobiotin to biotin (see Figure 1).
By measuring the amount of biotin in the new cells, and recognizing the 2:1 stoichiometric relationship between reduced electron donors and biotin molecules, the amount of electron donors present and used can be indirectly measured. Results showed that __________, as seen below:
